PT  - JOURNAL ARTICLE
AU  - Meta Heidenreich
AU  - Joseph M. Georgeson
AU  - Emanuele Locatelli
AU  - Lorenzo Rovigatti
AU  - Saroj Kumar Nandi
AU  - Avital Steinberg
AU  - Yotam Nadav
AU  - Eyal Shimoni
AU  - Samuel A. Safran
AU  - Jonathan P. K. Doye
AU  - Emmanuel D. Levy
TI  - Designer protein assemblies with tunable phase diagrams in living cells
AID  - 10.1101/2020.06.03.131433
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.06.03.131433
4099  - http://biorxiv.org/content/early/2020/06/03/2020.06.03.131433.short
4100  - http://biorxiv.org/content/early/2020/06/03/2020.06.03.131433.full
AB  - The self-organization of proteins into specific assemblies is a hallmark of biological systems. Principles governing protein-protein interactions have long been known. However, principles by which such nanoscale interactions generate diverse phenotypes of mesoscale assemblies, including phase-separated compartments, remains challenging to characterize and understand. To illuminate such principles, we create a system of two proteins designed to interact and form mesh-like assemblies in living cells. We devise a novel strategy to map high-resolution phase diagrams in vivo, which provide mesoscale self-assembly signatures of our system. The structural modularity of the two protein components allows straightforward modification of their molecular properties, enabling us to characterize how point mutations that change their interaction affinity impact the phase diagram and material state of the assemblies in vivo. Both, the phase diagrams and their dependence on interaction affinity were captured by theory and simulations, including out-of-equilibrium effects seen in growing cells. Applying our system to interrogate biological mechanisms of self-assembly, we find that co-translational protein binding suffices to recruit an mRNA to the designed micron-scale structures.Competing Interest StatementThe authors have declared no competing interest.